Sigenome brca2 smartpool

Cells were transfected with siRNAs using Dharmafect1 (Horizon Discovery) or INTERFERin (Polyplus) according to manufacturer’s instructions. Briefly, cells were transfected with 40 nM siRNAs by reverse transfection. The next day, the medium was replaced by fresh medium. The following siRNAs were used: siBRCA2 (Qiagen, SI02653434), siGENOME BRCA2 SMARTpool (Dharmacon, M-003462-01), siTP53 (Dharmacon, GCA UCU UAU CCG AGU GGA AUU UU), siRAD52 SMARTpool (Dharmacon, L-011760-00-0005). AllStars Negative Control siRNA (Qiagen, 1027281) or ON-TARGETplus non-targeting pool siRNA (Dharmacon, D-001810-10-05) were used as control.

Cells were transfected with siRNA using Dharmafect 1 (Dharmacon). Briefly, 0.25–2 × 10⁶ cells were transfected with 40 nM siRNA by reverse transfection. Following incubation for 24 or 48 h, depletion was determined by immunoblotting. Sequences of siRNAs used were as follows: siβ-catenin 5′-AUC AAC UGG AUA GUC AGC ACC-3′ (Dharmacon), siBRCA1 5′-CAG CAG UUU AUU ACU CAC UAA-3′ (Qiagen), siGENOME BRCA2 SMARTpool (Dharmacon, M-003462-01), siCDKN1A (p21, Dharmacon, J-003471-11), siCDKN1A (p21, Ambion, s416), siGSK3α (Thermo Fisher Scientific, 4390824; s6238), siGSK3β (Thermo Fisher Scientific, 4390824; s6241), siGSK3α/β (Cell Signaling, 6301), siMYC SMARTpool (Dharmacon, L-003282-02). AllStars siRNA (Qiagen, 1027281) was used as a negative control.